Part:BBa_K1777008

RNA accelerating cyclization

This RNA can bind to the flanking region of exon4 of hsACTB in HEK293T cells and facilitate the back-splicing mechanism. This part can be used to cyclize certain RNA exon.

Fuction

We design this RNA to promote the cyclization of hsACTB in HEK293T cells.

Results

Forty-eight hours after transfection with pZW1-sno-ACTB into HEK 293T cells, we use the specific circular exon-4 primers to test the circular RNA.

Figure 1. Circular RNA form with acRNA (pZW1-sno-ACTB).
HEK-293T cells were seeded into a 24-well plate. Forty-eight hours post-transfection, the total RNAs are extracted. Use circular RNA specific primers (have been BLAST on UCSC-In Silico, proved to be specific) doing PCR analysis, Unexpectedly, we amplify other circular RNAs. Extract non-specific gel and sequence it.Both vector without insert pZW1-sno and plasmid with acRNA pZW1-sno-ACTB appear non-specific electrophoretic band.
After sequencing, we found 200bp: circular RNA consists of exon4-exon3-exon4, 500bp: circular RNA onsists of exon4-exon5-exon6-exon2-exon3-exon4 while 700bp: circular RNA consists of exon4-exon5-exon3-exon4.